Application of histone transmethylase gene in regulating and controlling rice flowering period and fringe type

A methyltransferase and histone technology, applied in the field of plant genetic engineering, can solve problems such as rare reports and achieve the effect of alleviating food shortages

Inactive Publication Date: 2013-12-04
HUAZHONG AGRI UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

This indicates that the structural characteristics of CLF itself have a cert...

Method used

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  • Application of histone transmethylase gene in regulating and controlling rice flowering period and fringe type
  • Application of histone transmethylase gene in regulating and controlling rice flowering period and fringe type
  • Application of histone transmethylase gene in regulating and controlling rice flowering period and fringe type

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Experimental program
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Effect test

Embodiment 1

[0037] Embodiment 1: Amplification of full-length cDNA of SDG711 gene

[0038] To the gene SDG711 (gene accession number LOC_Os06g16390) needed by the present invention, mainly by RT-PCR method (referring to: J. Sam Brook, EF Fritsch, T Mani Artis works, Huang Peitang, Wang Jiaxi etc. translation, molecular cloning Experiment Guide (Third Edition), Beijing, Science Press, 2002 Edition) was amplified to obtain three sequences of the SDG711 gene, and then obtained the full length of the gene by restriction enzyme digestion. The specific operation is as follows:

[0039] 1) RNA was extracted from the leaves of the japonica rice variety Zhonghua 11 (ZH11) (a gift from researcher Li Meifang of the Institute of Crop Science, Chinese Academy of Agricultural Sciences, and it is a rice variety publicly used in China). The RNA extraction reagent was Trizol from Invitrogen. Extraction kit (see the instruction manual of the kit for specific operation steps);

[0040] 2) The steps to syn...

Embodiment 2

[0053] Example 2: Construction of SDG711 double-strand suppression vector

[0054] To the gene needed by the present invention, by RT-PCR method (referring to: J. Sam Brook, EF Fritsch, T Mani Atis is written, Huang Peitang, Wang Jiaxi etc. are translated, molecular cloning experiment guideline (third edition), Science Press, 2002 Edition) to amplify to obtain a specific sequence of SDG711, the specific steps are: according to the full-length cDNA sequence of the SDG711 gene announced by the public database (http: / / rice.plantbiologymsu.edu / ), through the ChromDB database (http: / / www.chromdb.org / index.html) to search for SDG711-specific sequences and design primers to amplify the RNAi inhibitory fragment by PCR. The amplified product was connected to pGEMT-vector (purchased from Promega (Beijing) Biotechnology Co., Ltd., ie Promega, USA) by T / A cloning for sequencing verification.

[0055] The primers used to clone the RNAi suppressor fragment of SDG711 are as follows:

[005...

Embodiment 3

[0064] Example 3: Transformation of binary Ti plasmid vector and detection of positive and expression levels of transgenic plants

[0065] 1) The newly constructed suppression expression vector pDS1301-SDG711-2 (from Example 2) was introduced into Agrobacterium EHA105 (purchased From the CAMBIA laboratory in Australia, which is a conventional commercial strain) strain. The transformed strain was named TR-SDG711.

[0066] 2) Transform the TR-SDG711 obtained in the previous step into the japonica rice variety Zhonghua 11 (ZH11). The transformation method refers to the method reported by Hiei et al. (Hiei et al., Efficient transformation of rice (Oryza sativa L.) mediated by Agrobacterium and sequence analysis of the boundaries of the T-DNA. Plant J, 1994, 6: 271-282.) and the standard method of the State Key Laboratory of Crop Genetic Improvement, Huazhong Agricultural University (for example: Patent No. ZL200710053552.9, the name of the invention: Rice Guangqin Separation, cl...

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Abstract

The invention belongs to the technical field of plant transgenic engineering, and particularly relates to application of histone transmethylase gene in regulating and controlling a rice flowering period and a fringe type. The histone transmethylase gene SDG711 is obtain by cloning, and the nucleotide sequence is shown in SEQ ID NO: 1. Through the method of double-strand RNA inhibition, transformation rice corresponding to the histone transmethylase gene is obtained, the phenomenon that the flowering time of the plant is brought forward for about 2 weeks under a long-day condition after the histone transmethylase gene is inhibited is discovered, and the fringe type of the plant is much smaller than that of a wild plant. A detection to the modification condition of the transgenic histone of the transgene shows that the methylation and trimethylation levels of the histone H3K27 of the transgenic plant are obviously changed. By regulating and controlling the rice histone methylated modification and affecting the flowering time and fringe size of rice, the histone transmethylase gene SDG711 can affect the output of the rice.

Description

technical field [0001] The invention belongs to the technical field of plant genetic engineering. It specifically relates to the isolation, cloning, functional verification and application of a histone methyltransferase gene SDG711 that affects the flowering period and panicle type of rice. Background technique [0002] Histone modification is one of the important mechanisms of epigenetic regulation, and H3K27me3 is a trimethylation modification that occurs on the 27th lysine of histone H3. At the level of gene expression regulation, H3K27me3 is often associated with transcriptional repression. The H3K27me3 transferase gene CLF in Arabidopsis is involved in the PcG complex-mediated silencing of floral organ homeotic genes PI and AP3, controlling the transition from vegetative growth to reproductive growth. It not only affects the morphogenesis of Arabidopsis flowers, but also plays a very important role in the regulatory pathway of flowering time. Studies have shown that ...

Claims

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Application Information

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IPC IPC(8): C12N15/54C12N9/10
Inventor 周道绣刘小云赵毓
Owner HUAZHONG AGRI UNIV
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